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目的探讨等位基因特异性聚合酶链反应(AS-PCR)检测单核苷酸多态性(SNP)的方法,研究鲑鱼精DNA对优化等位基因特异性引物的作用。方法对鲑鱼精DNA设一系列浓度梯度,分别进行AS-PCR后观察电泳结果。结果鲑鱼精DNA能显著提高识别基因SNP位点的特异性,在25μl的质粒检测系统中加入50 ng的鲑鱼精DNA结果最佳。结论鲑鱼精DNA加入到质粒检测系统中可有效测试所设计的检测引物的特异性,达到模拟基因组DNA的目的 。
Objective To investigate the method of allele-specific polymerase chain reaction (AS-PCR) for detecting single nucleotide polymorphisms (SNPs) and to study the effect of salmon sperm DNA on optimizing allele-specific primers. Methods A series of concentration gradients were set up for salmon sperm DNA. The results of electrophoresis were observed after AS-PCR respectively. Results Salmon sperm DNA significantly increased the specificity of SNP loci for recognition of the gene. The best results were obtained with 50 ng of salmon sperm DNA in a 25 μl plasmid detection system. Conclusion The addition of salmon sperm DNA into the plasmid detection system can effectively test the specificity of the designed detection primers and achieve the purpose of mimicking genomic DNA.